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różnorodność peptydów przeciwdrobnoustrojowych bezkręgowców

100%
Zdybicka-Barabas A., Cytryńska M.
Kosmos
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2017
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vol. 66
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issue 4
563-574
PL
Peptydy przeciwdrobnoustrojowe są kluczowymi efektorami odporności wrodzonej. Wykazują działanie przeciwbakteryjne, przeciwgrzybowe, przeciwpierwotniacze, a często przeciwwirusowe i przeciwnowotworowe. Wiele z nich zaangażowanych jest w neutralizację endotoksyn patogenów oraz ma właściwości immunomodulacyjne, stąd określane są również jako peptydy odpornościowe. O bogactwie i różnorodności naturalnie występujących peptydów przeciwdrobnoustrojowych świadczy ich liczba zgromadzona w bazie Antimicrobial Peptide Database (aps.unmc.edu/AP/main.php), która zawiera dane dotyczące ponad 2100 peptydów zidentyfikowanych u zwierząt. Spośród nich ponad 570 to peptydy odpornościowe bezkręgowców, w tym 495 opisanych u stawonogów. Niezwykle szerokie występowanie peptydów przeciwdrobnoustrojowych u przedstawicieli wszystkich królestw jednoznacznie wskazuje na ich fundamentalną rolę w sukcesie ewolucyjnym złożonych organizmów wielokomórkowych. Ich zasadnicze znaczenie w odporności bezkręgowców podkreśla ponadto fakt, że większość opisanych pod tym względem gatunków wytwarza zestaw peptydów zróżnicowanych pod względem struktury przestrzennej, właściwości biochemicznych, mechanizmu działania oraz spektrum aktywności przeciwdrobnoustrojowej.
EN
Antimicrobial peptides (AMPs) are the key effectors of innate immunity. They exhibit antimicrobial, antifungal, antiprotozoal, and often antiviral and anticancer activities. Many of them are involved in neutralization of pathogen endotoxins, have immunomodulatory properties, and are therefore referred to as defense peptides (host defense peptides). The wealth and diversity of naturally occurring AMPs is evidenced by their numbers in the Antimicrobial Peptide Database (aps.unmc.edu/AP/main.php), which contains data on over 2100 peptides identified in animals. Of these, over 570 are invertebrate peptides, including 495 described in arthropods. The unusually widespread presence of AMPs in all kingdoms clearly indicates their fundamental role in the evolutionary success of complex multicellular organisms. Their essential role in invertebrate immunity further emphasizes the fact that most species produce a set of defense peptides varied in terms of spatial structure, biochemical properties, mechanism of action and spectrum of antimicrobial activity.
2
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Phosphorylation of yeast ribosomal proteins by CKI and CKII in the presence of heparin

81%
Wojda I., Cytryńska M., Frajnt M., Jakubowicz T.
Acta Biochimica Polonica
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1999
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vol. 46
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issue 1
211-215
3
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Three Pseudomonas aeruginosa strains with different protease profiles

81%
Andrejko M., Zdybicka-Barabas A., Janczarek M., Cytryńska M.
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2013
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vol. 60
|
issue 1
83-90
EN
The proteolytic activity of three Pseudomonas aeruginosa strains, ATCC 27853 - a reference strain, and two clinical isolates was tested. The activity was examined after culturing the bacteria in two different growth media: the minimal M9 medium and rich Luria-Bertani broth (LB). Based on zymograms and protease activity specific assays, it was concluded that the reference strain produced three proteolytic enzymes in the LB medium: protease IV, elastase B and elastase A, while alkaline protease was only produced in the M9 medium. The clinical isolates of P. aeruginosa produced elastase B and alkaline protease when grown in the LB medium and the minimal M9 medium, respectively. PCR analysis confirmed the presence of both the lasB gene encoding elastase B and aprA coding for alkaline protease in the genomes of the three P. aeruginosa strains analyzed. The expression of these genes coding for two important P. aeruginosa virulence factors was dependent on the growth conditions in all the strains studied. The contribution of the extracellular proteinases to the virulence of P. aeruginosa strains used in this study was investigated using an insect model, the greater wax moth Galleria mellonella.
4
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The effect of vanadate on Pichia pastoris growth, protein kinase A activity and ribosomal protein phosphorylation.

81%
Frajnt M., Cytryńska M., Jakubowicz T.
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2002
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vol. 49
|
issue 4
959-968
EN
It was found that wild type yeast Pichia pastoris can tolerate vanadate concentration as high as 25 mM in the growth medium. Moreover, four vanadate-resistant P. pastoris strains designated JC100/1, JC100/3, JC100/9 and JC100/15 exhibiting tolerance up to 150 mM vanadate were selected. Growth of P. pastoris was correlated with vanadate to vanadyl reduction and its accumulation in the growth medium. In two selected strains, JC100/9 and JC100/15, protein kinase A activity was much higher in comparison to the wild type strain even without vanadate addition to the growth medium. Moreover, in the presence of vanadate, protein kinase A activity was significantly increased in the wild type and the vanadate-resistant JC100/1 and JC100/3 strains. It was also found that phosphorylation of a 40 kDa protein associated with ribosomes occured in all vanadate-resistant strains from the logarithmic, while in the wild type strain from the stationary growth phase. From the presented results it can be concluded that a protein kinase A signalling pathway(s) might be involved in the mechanism of P. pastoris vanadate resistance. The results also indicate a possible role of the 40 kDa protein in protection of P. pastoris against vanadate toxicity.
5
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Phosphorylation of acidic ribosomal proteins by ribosome-associated protein kinases of Saccharomyces cerevisiae and Schizosaccharofnyces pombe

81%
Jakubowicz T., Cytryńska M., Kowalczyk W., Gąsior E.
|
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vol. 40
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issue 4
497-505
6
Content available remote

The involvement of protein kinase A in the immune response of Galleria mellonella larvae to bacteria

81%
Cytryńska M., Zdybicka-Barabas A., Jakubowicz T.
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2007
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vol. 54
|
issue 1
167-174
EN
The role of protein kinase A (PKA) in the humoral immune response of the greater wax moth Galleria mellonella larvae to live Gram-positive bacteria Micrococcus lysodeikticus and Gram-negative bacteria Escherichia coli was investigated. The immune challenge of larvae with both kinds of bacteria caused an increase in fat body PKA activity depending on the injected bacteria. Gram-positive M. lysodeikticus was a much better inducer of the enzyme activity than Gram-negative E. coli. The PKA activity was increased about 2.5-fold and 1.5-fold, after M. lysodeikticus and E. coli injection, respectively. The in vivo inhibition of the enzyme activity by a cell permeable selective PKA inhibitor, Rp-8-Br-cAMPS, was correlated with considerable changes of fat body lysozyme content and hemolymph antimicrobial activity in bacteria-challenged insects. The kinetics of changes were different and dependent on the bacteria used for the immune challenge of G. mellonella larvae.
7
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The effect of cAMP and cGMP on the activity and substrate specificity of protein kinase A from methylotrophic yeast Pichia pastoris.

81%
Frajnt M., Cytryńska M., Jakubowicz T.
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2003
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vol. 50
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issue 4
1111-1118
EN
Cyclic AMP dependent protein kinase (PKA) from Pichia pastoris yeast cells was found to be activated by either cAMP or cGMP. Analogs of cAMP such as 8-chloro-cAMP and 8-bromo-cAMP were as potent as cAMP in PKA activation while N6,2'-O-dibutyryl-cAMP did not stimulate the enzyme activity. It was shown that protamine sulfate was almost equally phosphorylated in the presence of 1-2 × 10-6 M cAMP or cGMP while other substrates such as Kemptide, ribosomal protein S6, were phosphorylated to a lower extent in the presence of cGMP. It was demonstrated that pyruvate kinase is a substrate of PKA which co-purified with the P. pastoris enzyme. Moreover, pyruvate kinase was phosphorylated by PKA in the presence of cAMP and cGMP to comparable levels.
8
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Protein kinases CKI and CKII are implicated in modification of ribosomal proteins of the yeast Trichosporon cutaneum

81%
Wojda I., Cytryńska M., Frajnt M., Jakubowicz T.
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2002
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vol. 49
|
issue 4
947-957
EN
Phosphorylation of acidic ribosomal proteins P1/P2-P0 is a common phenomenon in eukaryotic organisms. It was found previously that in Trichosporon cutaneum, unlike in other yeast species, in addition to the two acidic ribosomal proteins, two other proteins of 15 kDa and 19 kDa of the small ribosomal subunit were phosphorylated. Here we describe two protein kinases: CKI and CKII, which are engaged in the modification of T. cutaneum ribosomal proteins. The acidic ribosomal proteins and the protein of 19 kDa were modified by CKII associated with ribosomes, while the protein of 15 kDa was modified by CKI. Protein kinase CKI was purified from cell-free extract (CKIC) and from ribosomal fraction (CKIR). The molecular mass of CKIC was established at 33 kDa while that of CKIR at 35-37 kDa. A protein of 40 kDa copurified with CKIR but not CKIC. Heparin significantly increased 40 kDa protein phosphorylation level by CKIR. Microsequencing analysis revealed the presence of CKI recognition motifs in the N-terminal fragment of the 40 kDa protein.
9
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The effect of Galleria mellonella hemolymph polypeptides on Legionella gormanii

71%
Chmiel E., Palusinska-Szysz M., Zdybicka-Barabas A., Cytryńska M., Mak P.
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2014
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vol. 61
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issue 1
123-127
EN
Among Legionella species, which are recognized to be pathogenic for humans, L. gormanii is the second prevalent causative agent of community-acquired pneumonia after L. pneumophila. Anti-L. gormanii activity of Galleria mellonella hemolymph extract and apolipophorin III (apoLp-III) was examined. The extract and apoLp-III at the concentration 0.025 mg/ml caused 75% and 10% decrease of the bacteria survival rate, respectively. The apoLp-III-induced changes of the bacteria cell surface were analyzed for the first time by atomic force microscopy. Our studies demonstrated the powerful anti-Legionella effects of the insect defence polypeptides, which could be exploited in drugs design against these pathogens.
10
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Phosphorylation of A-proteins by protein kinases bound to east ribosomes

70%
Cytryńska M., Jakubowicz T., Gąsior E.
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